Johnson, R. C. (Fort Detrick, Frederick, Md.) and N. D. Gary. Nutrition of Leptospira pomona. II. Fatty acid requirements. J. Bacteriol. 85:976-982. 1963.-The albumin fraction of rabbit serum, obtained by (NH(4))(2)SO(4) fractionation, supported good growth of Leptospira pomona Wickard when added to a medium containing phosphate buffer, NH(4)Cl, and thiamine. Extraction of the albumin fraction with ethanol and ether (3:1) resulted in a loss of its growth-supporting activity, which could be restored upon the addition of the extract or various fatty acids. The growth-supporting activity of fatty acids containing 2 to 18 carbon atoms was investigated with the extracted albumin medium. The activity of the fatty acid was found to be related to the number of carbon atoms in the molecule. Maximal growth was obtained with palmitic, heptadecanoic, stearic, and oleic acids. The amount of growth of L. pomona increased at a fixed concentration of albumin as the molecular ratio of extracted albumin to the long-chain fatty acids increased from 1:1 to 1:6. At higher ratios, growth decreased sharply. Direct utilization of fatty acids during growth of L. pomona was shown by the incorporation of palmitic acid-1-C(14) or -2-C(14) into cellular material. A medium composed of 0.02 m Na(2)HPO(4)-KH(2)PO(4) buffer (pH 7.4), 0.8% extracted albumin, 4 x 10(-4)m palmitic acid, 10(-3)m NH(4)Cl, and 5 mug/ml of thiamine supported good growth. L. pomona was transferred ten times, and seven other serotypes were transferred five times, in this medium without any decrease in the amount of growth. Growth of L. pomona was initiated with approximately four organisms per ml.